E53 E54 P13 P14 P15 P16 E8 E9 E10 E11 E12 E13 E14

Procedure

• Primer info

Samples E53 E54 P13 P14 P15 P16 E8 E9 E10 E11 E12 E13 E14 sed in this PCR (polimerase chain reaction) reaction

First A Master Mix of water, 2x mytaq mix, CA6.38R Primer (concentration: 10 micromolar), CA6.38L Primer (concentration: 10 micromolar)

Component	Volume for 14 reactions(+5% error)
Water	132.3 micoleters
2x mytaq mix	183.75 micoleters
CA6.38R Primer (concentration: 10 micromolar)	14.7 micoleters
CA6.38L Primer (concentration: 10 micromolar)	14.7 micoleters
BAS (Borine Serum Albumin)	1 microliter

24 micoleters of this master mix was added to 14 tubes For the 2 negative controls 12.5 micoleters water and 12.5 micoleters mytaq mix were added there was no positive control was available

1 microleter of the DNA samples was added to the coresponding tube

This PCR was set to Previous PCRs were showing up smeared and unclear. This could be becuase of too much amplified DNA

Step	Temperature	Duration	cycles
1	94°C	3:00	1
2	94°C	2:00	1
3	58.2°C	1:30	1
4	72°C	2:00	1
5	94°C	0:30	30
6	58.2°C	1:00	30
7	72°C	0:15	30
8	72°C	2:00	1
9	4°C	hold	-

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these amplified DNA samples were then tested by making a gel

to create the Gel the following steps were taken

2g agarose and 100ml 1X TAE buffer were mixed to make a 2% agarose 100ml gel mix the mix was microwaved and swirled until there were no flakes then 1 microliters Gelgreen was added

the mix was left to cool for 5 minutes then poured into the gel mold the gel was left to harden for 30 minutes

once hardened enough Used TAE buffer was poured into the gel box to cover the mold

5 microliter aliquots of DNA samples E53 E54 P13 P14 P15 P16 E8 E9 E10 E11 E12 E13 E14 two controls were made 1 microliter of 5x loading dye was added to each

First 3 microliters of the HyperLadder 100bp was added to the first well of the mold then the DNA samples were each added to the wells in the following order

Ladder	E53	E54	P13	Negative Control	P14	P15	P16	Positive control - E7
Ladder	E8	E9	E10	Negative Control	E11	E12	E13	E14

The voltage was set to 100V and run for 60 minutes

This is the resulting Gel

Notes

This Gel did not come out as well as some of the others, the PCR may have been messed up in some way.